ANTISENSE NUCLEIC ACIDS FOR THE PREVENTION AND TREATMENT OF DISORDERS IN WHICH EXPRESSION OF c-erbB PLAYS A ROLE

Abstract: The present invention is related to an antisense-nucleic acid or effective derivatives thereof hybridizing with an area of the messenger RNA (mRNA) or the DNA, encoding the p185<erbB-2> receptor (also termed c-erbB-2, HER2 or neu), a pharmaceutical composition, comprising an antisense nucleic acid or effective derivatives thereof hybridizing with an area of the messenger RNA (mRNA) or the DNA, encoding the c-erbB-2 receptor as well as the use of said antisense nucleic acids and derivatives thereof for the manufacturing of a pharmaceutical composition for the treatment of neoplasms and/or immune diseases and/or diseases involving pathological angiogenesis.

Description: Antisense nucleic Acids for the prevention and treatment of disorders in which expression of c-erbB plays a role The present invention is related to an antisense-nucleic acid or effective derivatives thereof hybridizing with an area of the messenger RNA (mRNA) or the DNA, encoding the pl85 erbB"2 receptor (also termed c-erbB-2, HER2 or neu) , a pharma¬ ceutical composition, comprising an antisense nucleic acid or effective derivatives thereof hybridizing with an area of the messenger RNA (mRNA) or the DNA, encoding the c-erbB-2 receptor as well as the use of said antisense nucleic acids and derivatives thereof for the manufacturing of a pharma¬ ceutical composition for the treatment of neoplasms and/or immune diseases and/or diseases involving pathological angiogenesis. ErbB-2 is a putative growth factor receptor with an intra- cellular tyrosine inase activity that is amplified and/or overexpressed by tumor cells in a variety of neoplasms inclu¬ ding breast cancer, lung cancer, esophageal and gastric can¬ cer, bile duct carcinoma, bladder cancer and ovarian cancer. In breast carcinoma patients, an amplification and overex- pression of the c-erbB-2 gene in the tumor tissue has been shown to correlate with a poor clinical prognosis. Overexpes- sion of pl85 erbB"2 in non-small-cell lung carcinoma has been shown to impart resistance to a number of chemotherapeutic agents . WO 93/09788 discloses a method for inhibiting the pro¬ liferation of cells which contain an erb B2/neu gene site. The method involves administering a therapeutic dose of an oligonucleotide which is capable of forming a colinear triplex with the promoter region of the erb B2/neu gene. WO 92/19732 discloses sense and antisense oligonucleotides, namely closed oligonucleotides. These compounds may be used pharmacologically as sense or antisense molecules. It is generally described the therapeutic use of oligonucleotides as sense or antisense agents. WO 92/13063 discloses a method for effecting expression of growth factors and growth factor receptors in cells or in multicellular animals and methods for testing compounds as effectors of transcription of growth factors and growth factor receptors . The article "Chemically Modified Oligodeoxynucleotide Analogs as Regulators of Viral and Cellular Gene Expression" in Gene Regulation: Biology of Antisense RNA and DNA discloses in general the use of chemically modified oligonucleotides in the antisense technology. It is an object of the present invention to provide a com¬ pound for the treatment of neoplasms and/or immune diseases and/or diseases involving pathological angiogenesis. The c-erbB-2 antisense-oligonucleotide of the invention solving the problem addressed above have the sequences as disclosed in the sequence listing under Seq. ID No. 1-105, having a DNA- or RNA-type structure. The control oligo¬ nucleotide has the sequence as disclosed in the sequence listing under Seq. ID No 106, having a DNA- or RNA-type structure . The antisense nucleic acids of the invention, were able to strongly inhibit the expression of the pl85 erbB~2 protein, tyrosine kinase activity and cell growth in a variety of tumor cells including breast cancer cells. Untransformed normal fibroblasts were not growth inhibited by the anti-c-erbB-2 antisense compounds. This suggests that pl85 erbB"2 plays a pathogenetic role in the growth of the above mentioned tumor cells. Furthermore, surprisingly, the immune response to a variety of neoplasms was significantly increased by the use of the antisense nucleic acids of the invention. Immune cell growth and activity was stimulated in co-culture assays culturing tumor cells and peripheral blood monocytes together. Surprisingly, the antisense nucleic acids of the invention, also acted as strong inhibitors of angiogenesis . This sug¬ gests, that either the secreted truncated form of the c-erbB-2 protein or the full receptor protein may play a causal role in pathological neoangiogenesis . According to the invention antisense nucleic acids or effec¬ tive derivatives thereof which hybridize with an area of the mRNA or DNA coding for pl85 erbB"2 can effectively treat the diseases addressed above. The antisense nucleic acid is able to hybridize with regions of pl85 erbB"2 mRNA. It is understood by the skilled person that fragments of the antisense nucleic acids and antisense nucleic acids containing these sequences work according to the invention so long as production of pl85 erbB"2 is reduced or inhibited. According to the invention the antisense-oligonucleotides are obtainable by solid phase synthesis using phosphite triester chemistry by growing the nucleotide chain in 3' -5' direction in that the respective nucleotide is coupled to the first nucleotide which is covalently attached to the solid phase comprising the steps of cleaving 5'DMT protecting group of the previous nucleotide, adding the respective nucleotide for chain propagation, modifying the phosphite group subsequently cap un- reacted 5' -hydroxyl groups and cleaving the oligonucleotide from the solid support, followed by working up the synthesis product. The chemical structures of oligodeoxy-ribonucleotides are given in figure 1 as well as the respective structures of antisense oligo-ribonucleotides are given in figure 2. The oligonucleotide chain is to be understood as a detail out of a longer nucleotide chain. In figure 1, lit. B means an organic base such as adenine (A) , guanine (G) , cytosine (C) and thymine (T) which are coupled via N9(A,G) or N1(D,T) to the desoxyribose. The sequence of the bases is the reverse complement of the genetic target sequence (mRNA-sequence) . The modifications used are 1. Oligodeoxy-ribonucleotides where all R are substituted by 1.1 R 1 = 0 1.2 R 1 = s 1.3 R 1 = F 1.4 R 1 = CH 3 1.5 R 1 = OEt 2. Oligodeoxy-ribonucleotides where R 1 is varied at the internucleotide phosphates within one oligonucleotide 5' B-p-B-p-B-p- (B-p-) B-p-B-p-B-p-B 3' I I I R la R la R la R ilb n R Ila R Ila R Ila where B = deoxy-ribonucleotide dA, dC, dG or dT de¬ pending on gene sequence p = internucleotide phosphate n = an oligodeoxy-ribonucleotide stretch of length 6 - 20 bases - H,.- 2.4 R la = CH 3 ; R lb = S 3. Oligodeoxy-ribonucleotides where R is alternated at the internucleotide phosphates within one oligo¬ nucleotide 5' B-p- (B-p-B-p) -B-p-B R la R lb R la R ib where B = deoxy-ribonucleotide dA, dC, dG or dT depending on gene sequence p = internucleotide phosphate n = an oligodeoxy-ribodinucleotide stretch of length 4 - 12 dinucleotides 3.2 R la = S; R lb = 0 3.2 R la = CH 3 ; R lb = 0 3.3 R la = S; R lb - CH 3 4. .Any of the compounds 1.1 - 1.5; 2.1 - 2.4; 3.1 - 3.3 2 coupled at R with the following compounds which are covalently coupled to increased cellular uptake 4.1 cholesterol 4.2 poly(L) lysine 4.3 transferrin 5. .Any of the compounds 1.1 - 1.5; 2.1 - 2.4; 3.1 - 3.3 coupled at R with the following compounds which are covalently coupled to increase cellular uptake 5.1 cholesterol 5.2 poly(L) lysine 5.3 transferrin In the case of the RNA-oligonucleotides (figure 2) are the basis (adenine (A) , guanine (G) , cytosine (C) , uracil (U) ) coupled via N9 (A,G) or Nl (C,U) to the ribose. The sequence of the basis is the reverse complement of the genetic target sequence (mRNA-sequence) . The modifications in the oligonu¬ cleotide sequence used are as follows 6. Oligo-ribonucleotides where all R are substituted by 6 . 1 R" = 0 6 . 2 R 1 = S 6 . 3 R 1 = F 6 . 4 R 1 = CH 3 6 . 5 R 1 = OEt Oligo-ribonucleotides where R is varied at the inter¬ nucleotide phosphates within one oligonucleotide 5' B-p-B-p-B-p- (B-p-) B-p-B-p-B-p-B 3' I I I R la R la R la R ilb n R Ila R Ila R Ila where B = ribonucleotide A, C, G or T depending on gene sequence p = internucleotide phosphate n = an oligo-ribonucleotide stretch of length 4 - 20 bases 7.1 R la = S; R lb = 0 7.2 R la = CH 3 ; R lb - 0 7.3 R la = S; R lb = CH., 7.4 R la = CH 3 ; R lb = S 8. Oligo-ribonucleotides where R is alternated at the in¬ ternucleotide phosphates within one oligonucleotide 5' B-p- (B-p-B-p) n-B-p-B 3' R la R lb R la R lb where B = ribonucleotide A, C, G or T depending on gene sequence p = internucleotide phosphate n = an oligo-ribodinucleotide stretch of length 4 -12 dinucleotides 8.2 R la = S; R lb - 0 8.2 R la = CH 3 ; R lb = 0 8.3 R la = S; R lb = CH 3 9. Any of the compounds 6.1 - 6.5; 7.1 - 7.4; 8.1 - 8.3 2 coupled at R with the following compounds which are covalently coupled to increase cellular uptake 9.1 cholesterol 9.2 poly(L) lysine 9.3 transferrin 10. .Any of the compounds 6.1 - 6.5; 7.1 - 7.4; 8.1 - 8.3 ^ the following compounds are covalently coupled to increased cellular uptake 10.1 cholesterol 10.2 poly(L) lysine 10.3 transferrin 11. Any of the compounds 6.1 - 6.5; 7.1 - 7.4; 8.1 - 8.3; 4 9.1 - 9.3; 10.1 - 10.3 where all R are substituted by 11 . 1 R' = 0 11 . 2 R 4 = F 11 . 3 R 4 = CH 3 In a preferred embodiment of the oligonucleotides of the in¬ vention they are phosphorothioate derivatives, having a DNA- or RNA-type structure. It is possible that one single individual sequence as mentioned above works as an antisense nucleic acid or oligo¬ nucleotide structure according to the invention. However, it is also possible that one strand of nucleotides comprises more than one of the sequences as mentioned above directly covalently linked or with other nucleotides covalently linked in between. Preferably, individual oligonucleotides are addressed. In a preferred embodiment of these oligo-nucleotides they are phosphorothioate derivatives. Modifications of the antisense-oligonucleotides are advantageous since they are not as fast destroyed by endoge¬ nous factors when applied as this is valid for naturally occurring nucleotide sequences. However, it is understood by the skilled person that also naturally occurring nucleot¬ ides having the disclosed sequence can be used according to the invention. In a very preferred embodiment the modifica¬ tion is a phosphorothioate modification. The synthesis of the oligodeoxy-nucleotide of the invention is described as an example in a greater detail as follows. Oligodeoxy-nucleotides were synthesized by stepwise 5'- addition of protected nucleosides using phosphite triester chemistry. The nucleotide A was introduced as 5'dimethoxy- trityl-deoxyadenosine (N-benzoyl) -N,N' -diisopropyl-2-cyano- ethyl phosphoramidite (0.1 M) ; C was introduced by a 5'- di ethoxytrityl-deoxycytidine(N 4-benzoyl) -N,N' -diisopropyl-2- cyanoethyl phosphoramidite; G was introduced as 5' -dimethoxy- Q trityl-deoxyguanosine (N -isobutyryl) -N,N' -diisopropyl-2- cyanoethyl phosphoramidite and the T was introduced as 5' - dimethodytrityl-deoxythymidine-N,N' -diisopropyl-2-cyanoethyl phosphoramidite. The nucleosides were preferably applied in 0.1 M concentration dissolved in acetonitrile. Synthesis was performed on controlled pore glass particles of approximately 150 μm diameter (pore diameter 500 A) to which the most 3' nucleoside is covalently attached via a long-chain alkylamine linker (average loading 30 μmol/g solid support) . The solid support was loaded into a cylindrical synthesis column, capped on both ends with filters which permit ade¬ quate flow of reagents but hold back the solid synthesis support. Reagents were delivered and withdrawn from the synthesis column using positive pressure of inert gas. The nucleotides were added to the growing oligonucleotide chain in 3'-> 5' direction. Each nucleotide was coupled using one round of the following synthesis cycle: Cleave 5'DMT (dimethoxytrityl) protecting group of the pre¬ vious nucleotide with 3-chloroacetic acid in dichloromethane followed by washing the column with anhydrous acetonitrile. Then simultaneously one of the bases in form of their protec¬ ted derivative depending on the sequence was added plus tetrazole in acetonitrile. After reaction the reaction mixture has been withdrawn and the phosphite was oxidized with a mixture of sulfur (So Q ) in carbon disulfide/pyridine/- triethylamine . After the oxidation reaction the mixture was withdrawn and the column was washed with acetonitrile. The unreacted 5' -hydroxyl groups were capped with simultaneous addition of 1-methylimidazole and acetic anhydride/lutidine- /tetrahydrofuran. Thereafter, the synthesis column was washed with acetonitrile and the next cycle was started. The work up procedure and purification of the synthesis pro¬ ducts occurred as follows. After the addition of the last nucleotide the deoxynucleo- tides were cleaved from the solid support by incubation in ammonia solution. Exocyclic base protecting groups were removed by further incubation in ammonia. Then the ammonia was evaporated under vacuum. Full-length synthesis products still bearing the 5'DMT protecting group were separated from shorter failure contaminants using reverse phase high per¬ formance liquid chromatography on silica C. 1o Q stationary phase. Eluents from the product peak were collected, dried under vacuum and the 5' -DMT protecting group cleaved by incubation in acetic acid which was evaporated thereafter under vacuum. The synthesis products were solubilized in the deionized water and extracted three times with diethylether. Then the products were dried in vacuo. Another HPLC-AX chromatography was performed and the eluents from the product peak were dialyzed against excess of Trisbuffer as well as a second dialysis against deionized water. The final products were lyophilized and stored dry. The antisense-nucleic acid of the invention can be used as pharmaceutical composition or medicament. This medicament can be used for treating neoplasms and/or immune diseases and/or diseases involving pathological angiogenesis in which the expression of c-erbB-2 derived receptor protein or truncated pl85 c"er B2 is of relevance for the pathogenicity. It can be used to reduce neoplastic cell growth in cells expressing pl85 c"erbB2 , to reverse resistance of tumor cells to the immune-response, to inhibit pathological angiogenesis and to stimulate the immune system. The antisense nucleic acids of the invention are intermediate products of the pharmaceutical composition or medicament of the invention. The pharmaceutical composition may comprise besides the effective compound(s) suitable carrier agents, solvents and other ingredients known in the art for producing medicaments. Preferably, these agents facilitate the administration of the pharmaceutical composition of the in¬ vention. Typically, the pharmaceutical composition is ad¬ ministered as i.v. infusion or i.v. bolus injection. The amount of the active ingredient to be administered is typically in the range of 0.2 - 50 mg of the oligonucleotide per kg body weight per day, in particular 1 - 12 mg/kg body weight per day. In principal the compound which can be used as an active com¬ pound in the pharmaceutical composition can be used as a diagnostic tool for evaluating whether the respective genes are expresses. Typically, radio active labelled nucleotides are hybridized by the method of northern blotting which is well-known in the art or in situ with a sample to be examined. The degree of hybridization is a measure for the degree of expression of the respective genes. The effect of c-erbB2 specific antisense-oligonucleotides on neoplastic cell growth was investigated. It was demonstrated that antisense oligodeoxynucleotides as well as phosphorothioate modified nucleic acids, complementary to c-erbB2 mRNA could specifically inhibit pl85 c"erbB2 protein expression and could to a significant amount reduce cell proliferation in breast cancer cells, ovarian carcinoma cells and bladder cancer cells. Also, it could be shown that protein synthesis and S6 kinase activity were strongly reduced in tumor cells, treated with the antisense nucleic acid. Furthermore, the immune response to a variety of neoplasms was significantly increased by the use of the antisense nucleic acids described below. Lymphocyte growth and activity was stimulated in co-culture assays culturing tumor cells and peripheral blood monocytes together. Furthermore, the antisense nucleic acids described above, also acted as inhibitors of angiogenesis. 94/04094 - 13 - SEQUENCE LISTING (1) GENERAL INFORMATION: (i) APPLICANT: (A) NAME: Biognostik Gesellschaft fuer biomolekulare Diagnostik mbH (B) STREET: Carl-Giesecke-Str. 3 (C) CITY: Goettingen (E) COUNTRY: Germany (F) POSTAL CODE (ZIP) : 37079 (ii) TITLE OF INVENTION: Antisense nucleic Acids for the prevention and treatment of disorders in which expression of c-erbB plays a role (iii) NUMBER OF SEQUENCES: 106 (iv) COMPUTER READABLE FORM: (A) MEDIUM TYPE: Floppy disc (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS (D) SOFTWARE: Patentin Release #1.0, Version #1.25 (EPO) (v) CURRENT APPLICATION DATA: APPLICATION NUMBER: EP 93120710.4 (2) INFORMATION FOR SEQUENCE ID NO: 1: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 1: TTCATGTCTG TGCC 14 (2) INFORMATION FOR SEQUENCE ID NO: 2: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 2: GTAGGTGAGT TCCA 14 (2) INFORMATION FOR SEQUENCE ID NO: 3: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 3: GTTGTGAGCG ATGA 14 (2) INFORMATION FOR SEQUENCE ID NO: 4: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 4: CATAGTTGTC CTCAAAGA 18 (2) INFORMATION FOR SEQUENCE ID NO: 5: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 5: GGCATAGTTG TCCT 14 (2) INFORMATION FOR SEQUENCE ID NO: 6: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 6: CATTGTCTAG CACG 14 (2) INFORMATION FOR SEQUENCE ID NO: 7: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 7: CTCCATTGTC TAGC 14 (2) INFORMATION FOR SEQUENCE ID NO: 8: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 8: GTATTGTTCA GCGG 14 (2) INFORMATION FOR SEQUENCE ID NO: 9: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 9: TCAAGATCTC TGTGAG 16 (2) INFORMATION FOR SEQUENCE ID NO: 10: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 10: CACAAAATCG TGTCCT 16 (2) INFORMATION FOR SEQUENCE ID NO: 11: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 11: TCCTTCCACA AAATCG 16 (2) INFORMATION FOR SEQUENCE ID NO: 12: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 12: GTGGAAGATG TCCT 14 (2) INFORMATION FOR SEQUENCE ID NO: 13: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 13: TCTTGTGGAA GATGTC 16 (2) INFORMATION FOR SEQUENCE ID NO: 14: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 14: TCTATCAGTG TGAGAG 16 (2) INFORMATION FOR SEQUENCE ID NO: 15: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 15: GGTTGGTGTC TATC 14 (2) INFORMATION FOR SEQUENCE ID NO: 16: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 16: ACATCGGAGA ACAG 14 (2) INFORMATION FOR SEQUENCE ID NO: 17: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 17: CCTTACACAT CGGA 14 (2) INFORMATION FOR SEQUENCE ID NO: 18: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 18: ACAATCCTCA GAACTC 16 (2) INFORMATION FOR SEQUENCE ID NO: 19: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 19: GCTCTGACAA TCCT 14 (2) INFORMATION FOR SEQUENCE ID NO: 20: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (Xi) SEQUENCE DESCRIPTION: SEQ ID NO: 20: TGGTTGAAGT GGAG 14 (2) INFORMATION FOR SEQUENCE ID NO: 21: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 21: CTGTGGTTGA AGTG 14 (2) INFORMATION FOR SEQUENCE ID NO: 22: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 22: GTTGTAGGTG ACCA 14 (2) INFORMATION FOR SEQUENCE ID NO: 23: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 23: CTGTGTTGTA GGTG 14 (2) INFORMATION FOR SEQUENCE ID NO: 24: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 24: GACTCAAACG TGTC 14 (2) INFORMATION FOR SEQUENCE ID NO: 25: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 25: CATGGACTCA AACG 14 (2) INFORMATION FOR SEQUENCE ID NO: 26: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 26: CGAATGTATA CCGG 14 (2) INFORMATION FOR SEQUENCE ID NO: 27: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 27: CCGAATGTAT ACCG 14 (2) INFORMATION FOR SEQUENCE ID NO: 28: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 28: GCCGAATGTA TACC 14 (2) INFORMATION FOR SEQUENCE ID NO: 29: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 29: GTAGTTGTAG GGAC 14 (2) INFORMATION FOR SEQUENCE ID NO: 30: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 30: TAGAAAGGTA GTTGTAGG 18 (2) INFORMATION FOR SEQUENCE ID NO: 31: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 31: GTAGAAAGGT AGTTGTAG 18 (2) INFORMATION FOR SEQUENCE ID NO: 32: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 32: CGTAGAAAGG TAGTTG 16 (2) INFORMATION FOR SEQUENCE ID NO: 33: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 33: CCGTAGAAAG GTAG 14 (2) INFORMATION FOR SEQUENCE ID NO: 34: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 34: GACCATAGCA CACT 14 (2) INFORMATION FOR SEQUENCE ID NO: 35: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 35: GGATATTGGC ACTG 14 (2) INFORMATION FOR SEQUENCE ID NO: 36: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 36: CCTGGATATT GGCA 14 (2) INFORMATION FOR SEQUENCE ID NO: 37: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 37: GCTCCCAAAG ATCT 14 (2) INFORMATION FOR SEQUENCE ID NO: 38: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 38: CCCATCAAAG CTCT 14 (2) INFORMATION FOR SEQUENCE ID NO: 39: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 39: CAAACACTTG GAGC 14 (2) INFORMATION FOR SEQUENCE ID NO: 40: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 40: GTCTCAAACA CTTGGA 16 (2) INFORMATION FOR SEQUENCE ID NO: 41 (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 41: GAGTCTCAAA CACTTG 16 (2) INFORMATION FOR SEQUENCE ID NO: 42: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 42: GTAACCTGTG ATCTCT 16 (2) INFORMATION FOR SEQUENCE ID NO: 43: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 43: GGTAACCTGT GATC 14 (2) INFORMATION FOR SEQUENCE ID NO: 44: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 44: GTATAGGTAA CCTGTG 16 (2) INFORMATION FOR SEQUENCE ID NO: 45: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 45: TGAGATGTAT AGGTAACC 18 (2) INFORMATION FOR SEQUENCE ID NO: 46: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 46: TGCTGAGATG TATAGG 16 (2) INFORMATION FOR SEQUENCE ID NO: 47: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 47: CCATGCTGAG ATGT 14 (2) INFORMATION FOR SEQUENCE ID NO: 48: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 48: GGATTACTTG CAGG 14 (2) INFORMATION FOR SEQUENCE ID NO:49: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 49: TGTTATGGTG GATGAG 16 (2) INFORMATION FOR SEQUENCE ID NO: 50: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 50: GGTGTTATGG TGGA 14 (2) INFORMATION FOR SEQUENCE ID NO: 51: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 51: GCAGTTGACA CACT 14 (2) INFORMATION FOR SEQUENCE ID NO: 52: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 52: AGTACTCGGC ATTC 14 (2) INFORMATION FOR SEQUENCE ID NO: 53: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 53: CATTCACATA CTCCCT 16 (2) INFORMATION FOR SEQUENCE ID NO: 54: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 54: TCCAAAACAG GTCACT 16 (2) INFORMATION FOR SEQUENCE ID NO: 55: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 55: GGTCCTTATA GTGG 14 (2) INFORMATION FOR SEQUENCE ID NO: 56: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 56: CAGAATGCCA ACCA 14 (2) INFORMATION FOR SEQUENCE ID NO: 57: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 57: ACGAGAATGC CAAC 14 (2) INFORMATION FOR SEQUENCE ID NO: 58: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 58: GATCCCAAAG ACCA 14 (2) INFORMATION FOR SEQUENCE ID NO: 59: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 59: TCGCTTGATG AGGA 14 (2) INFORMATION FOR SEQUENCE ID NO: 60: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 60: CATCGTGTAC TTCC 14 (2) INFORMATION FOR SEQUENCE ID NO: 61: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 61: GCATCGTGTA CTTC 14 (2) INFORMATION FOR SEQUENCE ID NO: 62: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 62: ACTGTGCCAA AAGC 14 (2) INFORMATION FOR SEQUENCE ID NO: 63: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 63: CTTGTAGACT GTGC 14 (2) INFORMATION FOR SEQUENCE ID NO: 64: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 64: CCCTTGTAGA CTGT 14 (2) INFORMATION FOR SEQUENCE ID NO: 65: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 65: TCAACACTTT GATGGC 16 (2) INFORMATION FOR SEQUENCE ID NO: 66: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 66: CCCTCAACAC TTTG 14 (2) INFORMATION FOR SEQUENCE ID NO: 67: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 67: GTGTTTTCCC TCAACA 16 (2) INFORMATION FOR SEQUENCE ID NO: 68: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 68: GTATGCTTCG TCTAAG 16 INFORMATION FOR SEQUENCE ID NO: 69 (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 69: CGTATGCTTC GTCT 14 (2) INFORMATION FOR SEQUENCE ID NO: 70: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid ( C ) STRA1-IDEDNESS : unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 70: CCATCACGTA TGCT 14 (2) INFORMATION FOR SEQUENCE ID NO: 71: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRA1TOEDNESS : unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 71: GCATAAGCTG TGTC 14 (2) INFORMATION FOR SEQUENCE ID NO: 72: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 72: CATGGTCTAA GAGG 14 (2) INFORMATION FOR SEQUENCE ID NO: 73: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 73: CAATCTGCAT ACACCA 16 (2) INFORMATION FOR SEQUENCE ID NO: 74: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 74: GGCAATCTGC ATAC 14 (2) INFORMATION FOR SEQUENCE ID NO: 75: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 75: CTGTCTCGTC AATG 14 (2) INFORMATION FOR SEQUENCE ID NO: 76: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS : unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 76: CATAACTCCA CACATC 16 (2) INFORMATION FOR SEQUENCE ID NO: 77: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 77: AGTCACACCA TAACTC 16 (2) INFORMATION FOR SEQUENCE ID NO: 78: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 78: ACAGTCACAC CATAAC 16 (2) INFORMATION FOR SEQUENCE ID NO: 79: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 79: CCCCAAAAGT CATC ■ 14 (2) INFORMATION FOR SEQUENCE ID NO: 80: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 80: TCGTAAGGTT TGGC 14 (2) INFORMATION FOR SEQUENCE ID NO: 81: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 81: GATCCCATCG TAAG 14 (2) INFORMATION FOR SEQUENCE ID NO: 82: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES ( i) SEQUENCE DESCRIPTION: SEQ ID NO: 82: CAATGGTGCA GATG 14 (2) INFORMATION FOR SEQUENCE ID NO: 83: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs . (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 83: GACATCAATG GTGC 14 (2) INFORMATION FOR SEQUENCE ID NO: 84: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 84: GTAGACATCA ATGGTG 16 (2) INFORMATION FOR SEQUENCE ID NO: 85: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 85: CATGATCATG TAGACATC 18 (2) INFORMATION FOR SEQUENCE ID NO: 86: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 86: CCATGATCAT GTAGAC 16 (2) INFORMATION FOR SEQUENCE ID NO: 87: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 87: CATTTGACCA TGATCATG 18 (2) INFORMATION FOR SEQUENCE ID NO: 88: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 88: CCAACATTTG ACCATG 16 (2) INFORMATION FOR SEQUENCE ID NO: 89: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 89: TCATCCAACA TTTGACCA 18 (2) INFORMATION FOR SEQUENCE ID NO: 90: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 90: GAGTCAATCA TCCAACAT 18 (2) INFORMATION FOR SEQUENCE ID NO: 91: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 91: CAGAGTCAAT CATCCA 16 (2) INFORMATION FOR SEQUENCE ID NO: 92: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 92: CCGACATTCA GAGT 14 (2) INFORMATION FOR SEQUENCE ID NO: 93: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 93: GAATTCAGAC ACCAAC 16 (2) INFORMATION FOR SEQUENCE ID NO: 94: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 94: GATGACCACA AAGC 14 (2) INFORMATION FOR SEQUENCE ID NO: 95: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 95: CCATCAAATA CATCGG 16 (2) INFORMATION FOR SEQUENCE ID NO: 96: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 96: TCACCATCAA ATACATCG 18 (2) INFORMATION FOR SEQUENCE ID NO: 97: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 97: CAACGTAGCC ATCA 14 (2) INFORMATION FOR SEQUENCE ID NO: 98: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 98: ACGTCTTTGA CGAC 14 (2) INFORMATION FOR SEQUENCE ID NO: 99: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 18 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 99: CAAAAACGTC TTTGACGA 18 (2) INFORMATION FOR SEQUENCE ID NO: 100: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 100: GGCAAAAACG TCTTTG 16 (2) INFORMATION FOR SEQUENCE ID NO: 101: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 101: CAAAGGCAAA AACGTC 16 (2) INFORMATION FOR SEQUENCE ID NO: 102: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 102: GTGTCAAGTA CTCG 14 (2) INFORMATION FOR SEQUENCE ID NO: 103: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 103: GTAATAGAGG TTGTCG 16 (2) INFORMATION FOR SEQUENCE ID NO: 104: (i) SEQUENCE CHARACTERISTICS: (A) LENGH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 104: CCCAGTAATA GAGG 14 (2) INFORMATION FOR SEQUENCE ID NO: 105: (i) SEQUENCE CHARACTERISTICS: (A) LENGH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 105: CATGGTGCTC ACTG 14 (2) INFORMATION FOR SEQUENCE ID NO: 106: (i) SEQUENCE CHARACTERISTICS: (A) LENGH: 14 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: unknown (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: DNA (genomic) (iii) ANTI-SENSE: YES (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 106: GTGCCTGTAC GTAC 14

Claims: C l a i s An antisense nucleic acid or an effective derivative therefrom which is capable of treating or preventing neoplasms, immune diseases and/or diseases involving pathological angiogenesis, hybridizing with an area of the messenger RNA (mRNA) and/or DNA encoding c-erbB-2, comprising the following sequences identified in the listing under Seq. ID No. 1 - 105, having DNA- or RNA-type structure. Antisense oligonucleotides of claim 1 wherein the oligonucleotides are modified oligonucleotides such as phosphorothioate derivatives . Antisense nucleic acid or -oligonucleotides according to any one of the claims 1 and/or 2 obtainable by solid phase synthesis using phosphite triester chemistry by growing the nucleotide chain in 3' -5' direction in that the respective nucleotide is coupled to the first nu¬ cleotide which is covalently attached to the solid phase comprising the steps of cleaving 5'DMT protecting group of the previous nucleotide, adding the respective nucleotide for chain propa¬ gation, modifying phosphite groups subsequently cap unre- acted 5'-hydroxyl groups and cleaving the oligonucleotide from the solid support, followed by working up the synthesis product. 4. A pharmaceutical composition comprising an effective amount of a compound of any one of the claims 1 to 3 for the prevention and treatment of neoplasms and/or immune diseases and/or diseases involving pathological angiogenesis . 5. Use of a compound according to any one of the claims 1 to 3 for the preparation of a pharmaceutical composi¬ tion for the prevention and treatment of neoplasms and/or immune diseases and/or diseases involving patho¬ logical angiogenesis. 6. Use of a compound according to any one of the claims 1 to 3 as diagnostic agent. 7. Method of treating or preventing neoplasms and/or im¬ mune diseases and diseases involving pathological an¬ giogenesis by administering an effective amount of the compound according to any one of the claims 1 to 3 or a pharmaceutical composition of claim 4 to a patient suffering from disorders related with the expression of c-erbB-2.

Inventors: BRYSCH WOLFGANG, SCHLINGENSIEPEN GEORG-F, SCHLINGENSIEPEN KARL-HERMANN, SCHLINGENSIEPEN REIMAR, BRYSCH, WOLFGANG, SCHLINGENSIEPEN, GEORG-F., SCHLINGENSIEPEN, KARL-HERMANN, SCHLINGENSIEPEN, REIMAR
Applicants: BIOGNOSTIK GES, BRYSCH WOLFGANG, SCHLINGENSIEPEN GEORG F, SCHLINGENSIEPEN KARL HERMANN, SCHLINGENSIEPEN REIMAR, BIOGNOSTIK GESELLSCHAFT FUER BIOMOLEKULARE DIAGNOST, BRYSCH, WOLFGANG, SCHLINGENSIEPEN, GEORG-F., SCHLINGENSIEPEN, KARL-HERMANN, SCHLINGENSIEPEN, REIMAR
Application year: 1994
Main classification: C12N15
IPC codes: A61K31, A61P9, A61P35, A61P37, C07H21, C12N15, C12Q1